钩藤碱对DAT基因敲除ADHD模型小鼠多巴胺能神经元代谢的影响

Effects of rhynchophylline on the metabolism of dopaminergic neurons in DAT-knockout mice

目的研究钩藤碱对多巴胺转运体(dopamine transporter,DAT)基因敲除注意力缺陷多动障碍(attention-deficit hyperactivity disorder,ADHD)模型小鼠多巴胺能神经元及其代谢产物的影响。方法体外ADHD模型小鼠及正常小鼠神经元细胞,分为4组:健康对照组、DAT模型组、DAT+钩藤碱组、DAT+宁动颗粒组,各组干预后0、1、3、6、12小时收集细胞及细胞培养液,并ELASA法检测细胞培养液中多巴胺(dopamin,DA)、多巴胺代谢产物高香草酸(homovanillic acid,HVA)、代谢酶单胺氧化酶(monoamine oxidase,MAO)及神经营养因子(brain derived neutrophil factor,BDNF)含量。结果 DAT模型组中,在时间上3、6小时作用多巴胺的代谢达到高峰;DA浓度与DAT模型组及健康对照组比较,DAT+钩藤碱组与DAT+宁动颗粒组培养液中含量均明显降低(P<0.05),HVA浓度、MAO浓度、BDNF浓度均显著增高,且DAT+钩藤碱组与DAT+宁动颗粒组间无显著性差异。结论钩藤碱可以通过调节MAO活性及BDNF活性促进多巴胺能神经元代谢。

Objective To explore the effect of rhynchophylline （Rhy） on the metabolism of dopa- mine for DAT-Kn rats dopaminergic neurons. Methods DAT-Kn rats＇ dopaminergic neurons were extrac- ted, separated and cultured and then divided into 4 groups： the control group, DAT-Kn model group, DAT-Kn ＋ Rhy group and DAT-Kn ＋NDG group. The cells and cell eullure medium of each group were respectively collected in processing after the completion of 0, 1, 3, 6, 12h after intervention. After immu- nofluorescence staining, the content of DA, HVA, MAO and BDNF in cell cultm＇e medium was detected by ELASA. Results In the DAT model group, metabolism of dopamin reached to the peak at 3, 6h. Compared with the model group, cell culture medium levels of DA of the DAT-kn＋Rhy group and DAT-kn ＋NDG groups were decreased（P〈0.05）. Meanwhile the cell cultm＇e medimn levels of HVA, MAO and BDNF of the DAT-kn＋Rhy group and DAT-kn＋NDG group groups were significantly increased（P〈0.05）. Conclusion Rhy can adjust activity of the MAO and BDNF to promote the metabolism of dopaininergic neurons.